Development of a Cell-Based High-Throughput Functional Screening Assay To Study Host-Virus (COVID-19) Interaction

尽管需要预防和治疗，但尚未批准用于高致病性严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的药物或疫苗。在控制或预防SARS-CoV-2传播的方案中，用中和抗体、肽阻断剂或针对病毒（刺突蛋白）和宿主膜蛋白（ACE 2和TMPRSS 2）的蛋白酶抑制剂阻断其进入细胞可能对对抗入侵病毒非常有效。不幸的是，由于与病毒复制有关的安全性问题，SARS-CoV-2被归类为BSL 3制剂，其操作需要高度封闭的实验室和训练有素的人员。为了应对当前的大流行，与合作伙伴Immune Biosolutions（一家专注于发现、工程和开发用于肿瘤学和传染病的人源化鸡抗体的生物技术公司）提交的这一合作研究项目的目标是，是开发一种安全和准确的SARS-CoV-2感染慢病毒模型，这将允许高通量筛选和验证COVID-1。BSL 2实验室中的19种创新疗法。 为此，我们建议：（1）将SARS-CoV-2刺突蛋白（SARS-CoV-2S），并通过监测eGFP阳性假病毒颗粒的允许细胞感染，实现基于荧光检测的功能测定，用于高通量筛选和定量抗病毒药物功效;（2）筛选肺、胃肠道和脑细胞类型对假病毒感染的易感性。 预计这种基于慢病毒细胞的功能平台将加速发现和验证新的解决方案，以对抗当前和未来的冠状病毒感染。

Despite the needs for prophylactic and therapeutic treatments, no drug or vaccine has yet been approved for the highly pathogenic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Among the options envisaged to control or prevent the spread of SARS-CoV-2, blocking its cell entry with neutralizing antibodies, peptide blockers or protease inhibitors targeting viral (spike protein) and host membrane proteins (ACE2 and TMPRSS2) may be extremely effective to combat the invading virus. Unfortunately, due to the safety concerns related to viral replication, SARS-CoV-2 is classified as a BSL3 agent and its manipulation requires high-containment laboratories and trained personnel. In response to the current pandemic, the goal of this collaborative research project submitted with the partner Immune Biosolutions, a biotech company focusing on the discovery, engineering and development of humanized chicken antibodies for oncology and infectious diseases, is to develop a safe and accurate lentiviral model of SARS-CoV-2 infection that will allow high-throughput screening and validation of COVID-19 innovative therapeutics in BSL2 laboratories. To this aim, we propose (1) to incorporate the SARS-CoV-2 spike protein (SARS-CoV-2 S) in lentiviral pseudovirions and implement a robust and reproducible fluorescence detection-based functional assay for high-throughput screening and quantification of antiviral drug efficacy through the monitoring of permissive cell infection by eGFP-positive pseudoviral particles; and (2) to screen lung, gastrointestinal and brain cell types for their susceptibility to pseudovirus infection. It is anticipated that this lentiviral cell-based functional platform will accelerate the discovery and validation of new solutions to fight against current and future coronavirus infections.